食管癌细胞株中肿瘤干细胞的分选及生物学鉴定南京医科大学.docVIP

食管癌细胞株中肿瘤干细胞样细胞培养分离及生物学鉴定 汪建林，孙志强，于静萍，孙苏平* (南京医科大学附属常州第二人民医院放疗科,江苏 常州 213003) [摘 要] 目的:探讨用无血清培养基方法从人食管癌细胞株KYSE-150、TE-1中分离富含肿瘤干细胞的细胞球并鉴定其生物学特性。方法：运用无血清培养基培养KYSE-150、TE-1细胞，观察细胞球的生成及在有血清培养基中分化情况，利用MTT法以及Transwell小室法研究食管癌细胞球的增殖情况和迁移能力，流式细胞仪检测分子标记CD44+、CD271+、CD44+CD271+的表达。结果：KYSE-150、TE-1细胞在无血清培养基培养条件下可以获得可稳定传代的细胞球，无血清培养基培养下KYSE-150细胞球CD44+、CD271+、CD44+CD271+细胞分别为(64.92±11.04)%、(28.27±6.79)%、 (24.07±5.39)%，均显著高于亲本细胞的(37.04±6.30)%、(3.69±0.49)%、(1.64±0.11)%（P值均＜0.05）。 TE-1细胞球CD44+、CD271+、CD44+CD271+细胞占(6.41±0.87)%，(2.58±0.17)%、(0.27±0.53)%，均显著高于亲本细胞的(1.87±0.18)%、(0.44±0.10)%、 (0.093±0.025)%（P值均＜0.05）。结论：应用无血清培养基可以从KYSE-150、TE-1细胞系中能分离出具有干细胞特性的食管癌细胞球，分子CD44、CD271可能是食管癌干细胞的标志。 [关键词] 食管癌；肿瘤干细胞；无血清培养；细胞球； The study of isolation and biological characteristics of cancer stem cells from esophageal cancer cell lines Wang Jianlin，Sun Zhiqiang，Yu Jingping，Sun Suping * （Department of Radiation Oncology，Changzhou Second Hospital-Affiliated Hospital of NJMU，Changzhou 213003，China） [Abstract] objective：To study on the cultivation method for cancer stem cells（CSCs）in esophageal cancer cell lines KYSE-150 and TE-1 with serum-free medium（SFM）and identify whether resulting spheroid cells display cancer stem cells characteristics. Methods：KYSE-150 and TE-1 cells was cultured in serum-free medium. The cells were observed to generate the spheres and differentiation in serum-supplemented medium. MTT assay and invasive assay were applied to examine cell invasive ability of both KYSE-150 and TE-1 spheres and parental cells. CD44+、CD271+、CD44+CD271+ expression of tumor spheroid cells was also analyzed by flow cytometry. Results：After SFM culture，KYSE-150 and TE-1 cells formed spheres，and could continuously passage in vitro. Cell spheres differentiation recurred when serum was supplemented in SFM. CD44+、CD271+、CD44+CD271+ in KYSE-150 cell were [(64.92±11.04)%、(28.27±6.79)%、 (24.07±5.39)%],which were significantly higher than that those in parent cell [(37.04±6.30)%、(3.69±0.49)%、(1.64±0.11)